title: Separation of long RNA by agarose-formaldehyde gel electrophoresis.
abstract: We describe a method to facilitate electrophoretic separation of high-molecular-weight RNA species, such as ribosomal RNAs and their precursors, on agarose-formaldehyde gels. Two alternative "pK-matched" buffer systems were substituted for the traditionally used Mops-based conductive medium. The key advantages include shortened run times, a 5-fold reduction in formaldehyde concentration, a significantly improved resolution of long RNAs, and consistency in separation. The new procedure has a streamlined work flow that helps to minimize errors and is broadly applicable to agarose gel electrophoresis of RNA samples and their subsequent analysis by Northern blotting.
authors.affiliation: Department of Cell Biology, Rowan University, School of Osteopathic Medicine, Stratford, NJ 08084, USA.
authors.forename: Farrah H
authors.initials: FH
authors.lastname: Mansour
authors.forename: Dimitri G
authors.initials: DG
authors.lastname: Pestov
mesh_terms: D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005557:Formaldehyde; D008970:Molecular Weight; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification*; D012685:Sepharose
chemical_list: D005557:Formaldehyde; D012313:RNA; D012685:Sepharose
keywords: Electrophoresis; Northern blotting; Pre-rRNA; RNA; rRNA biogenesis